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Original Articles

Proximal Enhancer of the Human Insulin Receptor Gene Binds the Transcription Factor Sp1

  1. Hui Chen,
  2. Gillian E Walker,
  3. Simeon I Taylor and
  4. Catherine McKeon
  1. Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health Bethesda, Maryland
  1. Address correspondence and reprint requests to Dr. Catherine McKeon, Metabolic Diseases and Gene Therapy Research Program, Westwood Building, Room 621, National Institutes of Health, Bethesda, MD 20892.
Diabetes 1994 Jul; 43(7): 884-889. https://doi.org/10.2337/diab.43.7.884
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Abstract

The insulin receptor is a growth regulator present on the surface of most cells that transmits a mitogenic signal in response to insulin. Thus, the gene for the insulin receptor is constitutively expressed at low levels in all cells. We characterize a constitutive enhancer element that is present in the proximal promoter of the human insulin receptor gene. We have localized the enhancer to a 26–base-pair (26-bp) sequence from –528 to –503. When this sequence is inserted into the proximal promoter, a three- to fourfold increase in promoter activity is observed, and when two copies are inserted, a five- to sixfold increase is seen. Electrophoretic mobility shift analysis demonstrates that nuclear factors binding to this sequence are found in many different cell types. At least two proteins with different specificities bind within this 26-bp sequence. The identity of the predominant binding protein is Sp1, because an oligonucleotide composed of an Sp1 consensus binding sequence can compete for several of the DNA-protein complexes. In addition, we demonstrate that purified Sp1 can bind to the 26-bp oligonucleotide and that this complex comigrates with a DNA-protein complex formed with a HeLa nuclear extract. Finally, an antibody to human Sp1 protein is able to bind to the enhancer DNA/HeLa protein complex and supershift this complex. These findings suggest that this sequence corresponds to a general element that may contribute to the ubiquitous expression of the human insulin receptor gene.

  • Received April 30, 1993.
  • Revision received March 17, 1994.
  • Accepted March 17, 1994.
  • Copyright © 1994 by the American Diabetes Association

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July 1994, 43(7)
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Proximal Enhancer of the Human Insulin Receptor Gene Binds the Transcription Factor Sp1
Hui Chen, Gillian E Walker, Simeon I Taylor, Catherine McKeon
Diabetes Jul 1994, 43 (7) 884-889; DOI: 10.2337/diab.43.7.884

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Proximal Enhancer of the Human Insulin Receptor Gene Binds the Transcription Factor Sp1
Hui Chen, Gillian E Walker, Simeon I Taylor, Catherine McKeon
Diabetes Jul 1994, 43 (7) 884-889; DOI: 10.2337/diab.43.7.884
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